SEQUENCE SHAPE AND SPECIFICITY OF ANTIBODIES

某些近交系小鼠的免疫反应是由 具有共同可变区结构(独特型)的抗体 用抗独特型试剂进行血清学检测。如此独一无二的 表征某些抗体特异性的决定因素是有用的 抗体多样性和抗体多样性研究中的结构和遗传标记 监管。A/J小鼠主要的交叉反应独特型(ID-CR) Rho-azophenylarsate(ARS)-蛋白质结合物免疫 可遗传并由生殖系基因片段的单一组合编码 (“规范”)。尽管Jerne提出ID和ID之间的交互作用 抗-ID通过识别ID决定簇来调节免疫反应， ARS系统中的证据已经积累，ID主导地位可能是 以抗原为导向筛选具有较高抗原性的体细胞突变体 亲和力来源于偏爱的(更具“适应性”)的V区生殖系基因 组合。ARS系统是通过以下方式检查监管的模型 确定增强抗原所涉及的抗体结构变化 在免疫反应期间暂时发生的亲和力(“亲和力” 成熟“)。基于高分辨率的X射线晶体结构 携带Ars结合ID-CR的体细胞突变抗体(36-71)和 由此推导出的生殖系三级结构，我们现在可以检查 Ars结合和独特型的详细结构相关性，与 体细胞突变和基因连接变异。我们将充分利用 基于位点的蛋白质工程方法学研究进展 突变和表达，与预测性计算机建模相结合 晶体结构与：1)体细胞突变与基因有关 连接多样性对半抗原结合部位精细结构的影响 比较生殖系和生殖系三级结构的几何构型 突变的抗体。2)设计新的突变以增加 亲和力。3)检查体内发现的高突变部位，以评估 无论它们是否与亲和力增强或改变的白痴有关。 4)设计新的Ars同源物的结合特异性，作为一种测量 规范V区结构的分化能力。5)地图 诱变独特异体及其与单抗反应性的测定 抗ID试剂，并通过测定其晶体结构 ID-反ID。利用抗体工程学进一步了解和 因此，修饰抗原-抗体的互补性对于设计是必要的。 针对药物、毒素、 荷尔蒙和细胞受体。

Immune responses in certain inbred mouse strains are dominated by antibodies which share common variable (V) region structures (idiotypes) detected serologically by anti-idiotypic reagents. Such idiotypic determinants characterizing certain antibody specificities are useful structural and genetic markers in studies of antibody diversity and regulation. The predominant cross reactive idiotype (Id-CR) in A/J mice immunized with rho-azophenylarsonate (Ars)-protein conjugates is heritable and encoded by a single combination of germline gene segments ("canonical"). Although Jerne proposed that interactions between Id and anti-Id regulate immune responses through recognition of Id determinants, evidence in the Ars system has accumulated that Id dominance may be due to antigen-driven selection of favorable somatic mutants with higher affinity derived from preferred (more "adaptable") V region germline gene combinations. The Ars system is a model for examining regulation by defining the antibody structural changes involved in enhanced antigen affinity occurring temporally during the immune response ("affinity maturation"). Based upon the high resolution X-ray crystal structure of a somatically mutated Ars-binding Id-CR bearing antibody (36-71) and the germline tertiary structure deduced therefrom, we can now examine the detailed structural correlates of Ars binding and idiotypy, as related to somatic mutation and gene junctional variation. We will capitalize on methodologic advances in protein engineering using site-specific mutagenesis and expression, in concert with predictive computer modelling and the crystal structures to: 1) relate somatic mutation and gene junctional diversity to fine structure of the hapten binding site geometry using comparisons of the tertiary structures of germline and mutated antibodies. 2) Engineer novel mutations designed to increase affinity. 3) Examine sites of hypermutation found in vivo to assess whether or not they relate to affinity enhancement or altered idiotopes. 4) Engineer new binding specificities to Ars homologues as a measure of the differentiative capacity of a canonical V region structure. 5) Map idiotopes by mutagenesis and measurement of reactivity with monoclonal anti-Id reagents, and by determination of the crystal structures of Id-anti-Id. The use of antibody engineering to further understand and thus modify antigen-antibody complementarity is necessary to the design of antibodies for therapeutic use in targeting to drugs, toxins, hormones, and cellular receptors.